Fig. 4

Expression of NSD2 in prostate cancer and metastases. a Immunostaining of Nsd2 and other markers on mouse primary tumors and metastases. Shown are representative H&E images and immunostaining for the indicated antibodies from non-metastatic NP mice and metastatic NPK mice (n = 4/group). Scale bars in the low power H&E images represent 100 microns, and all other panels 50 microns. b, c Violin plots comparing mRNA expression levels of NSD2 in TCGA and SU2C human prostate cancer cohorts (Supplementary Table 1). b compares primary tumors from TCGA divided based on pathological grade [Gleason <4 (n = 104) or ≥ 4 (n = 173)], as indicated. c compares primary tumors and metastases from a cohort combining primary tumors from TCGA (all Gleason scores; n = 333) and metastases from SU2C (n = 51). P-values were estimated using the two-sample two-tailed Welch t-test. d, e Immunostaining of NSD2 on a human prostate tissue microarray (TMA) (n = 100 independent cases). d shows representative images representing benign prostate, untreated localized adenocarcinoma, castration-resistant adenocarcinoma (CRPC-Adeno) and neuroendocrine prostate cancer (NEPC). Nuclear staining intensity was evaluated blinded by a pathologist and scored as negative (or present in <5% of nuclei), weak, moderate or strong. Scale bars represent 50 microns. e shows quantification of nuclear intensity staining for each score (negative, weak, moderate, and strong). The p-values compare negative/weak staining versus moderate/strong staining in each group and were calculated using a two-tailed Fisher's exact test. f Immunostaining of NSD2 on matched patient sets of primary prostate cancer and distant metastasis to soft tissues or bone, as indicated. Patient 1 shows representative images of lower pathological grade (Gleason 3 + 3), which is negative for NSD2, and higher pathological grade (Gleason 4 + 5) and a liver metastasis that have increasing expression of NSD2. Patient 2 shows a high grade primary tumor (primary) that is negative for NSD2 and a matched bone metastasis in which NSD2 staining is readily detected. Scale bars represent 50 μ